The aim of the study was to estimate the diagnostic sensitivity (DSe) and specificity (DSp) of an agar gel immunodiffusion (AGID) assay for detection of myxoma virus (MYXV) in the classical form of myxomatosis and to compare its diagnostic performance to that of molecular methods (IAC-PCR, OIE PCR, and OIE real-time PCR). A panel of MYXV-positive samples of tissue homogenates with low (1 PCR unit – PCRU) and high (3,125 PCRU) virus levels and outbreak samples were used for method comparison studies. The validation parameters of the AGID assay were assessed using statistical methods. The AGID attained DSe of 0.65 (CI₉₅%: 0.53–0.76), DSp of 1.00 (CI₉₅%: 0.40–1.00), and accuracy of 0.67 (CI₉₅%: 0.55–0.76). The assay confirmed its diagnostic usefulness primarily for testing samples containing ≥3,125 PCRU of MYXV DNA. However, in the assaying of samples containing <3,125 PCRU of the virus there was a higher probability of getting false negative results, and only molecular methods showed a 100% sensitivity for samples with low (1 PCRU) virus concentration. The overall concordance of the results between AGID and IAC-PCR was fair (ĸ = 0.40). Full concordance of the results was observed for OIE PCR and OIE real-time PCR when control reference material was analysed. Findings from this study suggest that AGID can be used with some limitations as a screening tool for detection of MYXV infections.

Early detection of pregnancy is vital for appropriate reproductive management programmes to facilitate the rapid re-insemination of non-pregnant females and reduce the calving interval. A barium chloride test was compared with a commercial progesterone ELISA to detect pregnancy in non-descriptive cows and investigate if it could be applied as an alternative to ELISA in the field. Blood and urine samples were collected from 74 cows with recorded insemination dates. The progesterone ELISA and barium chloride assay were implemented to detect progesterone (P4) in blood and urine specimens, respectively. The cows' reproductive systems were examined after they were slaughtered to determine the uterus's status. Macroscopic examination of the uterus was used as a reference standard for both tests. The sensitivity rates of the P4 ELISA and barium chloride test to detect pregnant cows were 100.0% and 79.4%, and to detect the corpus luteum (CL) were 83.0% and 87.0%, respectively, their sensitivity increasing in the presence of the CL. The ELISA and barium chloride tests were 79.7% and 52.7% accurate in the diagnosis of pregnancy. The accuracy of the barium chloride test in CL detection increased to 81.0%, and that of the ELISA to 86.4%. There were no significant differences (P = 0.052) between the barium chloride assay and ELISA when they were utilised for the identification of the CL. The barium chloride test can be an inexpensive and time-saving alternative to ELISA in pregnancy diagnosis when the insemination date is known.

Objective: The objective of this study was to develop a low-cost kit for the detection of subclinical mastitis (SCM) and to check its validity, reproducibility, and efficacy at the field level. Materials and Methods: A total of 550 quarter milk samples from crossbred dairy cows were collected, of which 400 milk samples were used to validate the newly developed BLRI mastitis test (BMT) kit to justify its efficacy as an individual test kit in detecting SCM based on somatic cell count (SCC) by direct microscopic count (DMC). The efficacy of the newly developed BMT was compared with the California Mastitis Test (CMT) kit. Another 150 milk samples were subjected to SCC determined by DMC and DCC (De Laval cell counter®) categorized by CMT and BMT scores. Results: A SCM test kit, namely, BMT kit was successfully developed in this study. The percentage accuracy of CMT and BMT were 76.75% and 75.75%; sensitivity 69.36% and 67.56%; specificity 85.95% and 85.85%; positive predictive value 86.03% and 85.71%; negative predictive value 69.23% and 68%, respectively. A p value of 0.001 was found for both CMT and BMT. However, CMT and BMT had no significant difference in sensitivity (p = 0.778). Average SCCs (cells/ml) determined by DCC and DMC, respectively, were mostly corresponded to the SCC ranges of both CMT and BMT scores. Conclusion: The newly developed BMT kit is an independent, cheap, farmer-friendly, first country made, and reliable SCM diagnostic test kit that can be used at field condition. [J Adv Vet Anim Res 2019; 6(3.000): 425-430]

Intraocular production of Toxoplasma gondii-specific antibody in cats has been estimated by comparing the ratio of T gondii-specific antibody in aqueous humor and serum with the ratio of total immunoglobulins in serum and aqueous humor (Goldmann-Witmer coefficient; aqueous antibody coefficient; C value). It has been proposed that in human beings, comparison of the ratio of T gondii-specific antibody in aqueous humor and serum with the ratio of antibodies against a nonocular pathogen in serum and aqueous humor is more accurate than methods using total immunoglobulin quantification. We developed an ELISA for detection of calicivirus-specific antibodies in the serum and aqueous humor of cats. By evaluating calicivirus-specific antibody concentrations in the aqueous humor of healthy and diseased cats, calicivirus was assessed as a nonintraocular pathogen. The ratio of T gondii-specific antibodies in the aqueous humor and serum and the ratio of calicivirus-specific antibodies in serum and aqueous humor were evaluated as a means of estimating intraocular T gondii-specific antibody production. A field strain of feline calicivirus was isolated, cultured, and purified. A calicivirus-specific IgG ELISA was developed for detection of feline calicivirus-specific IgG in serum and aqueous humor. Calicivirus-specific IgG was measured in the serum and aqueous humor from 3 groups of control cats. Results suggested that calicivirus is a nonintraocular pathogen in cats and that calicivirus IgG detected in aqueous humor is attributable to leakage across a damaged blood-ocular barrier. Intraocular production of T gondii-specific antibodies was estimated, using 2 formulas. The C value was calculated by multiplying the ratio of T gondii-specific IgM or IgG in aqueous humor and serum by the ratio of total immunoglobulins (using the corresponding IgM or IgG class) in serum and aqueous humor. The Ctc value (Toxoplasma-calicivirus Goldmann-Witmer coefficient) was calculated by multiplying the ratio of T gondii-specific IgM or IgG in aqueous humor and serum by the ratio of calicivirus-specific IgG in serum and aqueous humor. Serum and aqueous humor samples were obtained from 41 client-owned cats with uveitis, and T gondii-specific C values and Ctc values were calculated. Toxoplasma gondii-specific IgM or IgG C values of 10 or greater or T gondii-specific IgM or IgG Ctc values of 1 or greater were considered to be suggestive of intraocular T gondii-specific antibody production. Of the 41 cats, 20 (48.7%) had evidence of intraocular production of T gondii-specific antibody on the basis of either an IgM or IgG C value of 10 or greater. A Ctc value could not be calculated in 3 cats because calicivirus-specific IgG was not present in aqueous humor. Of the 38 cats for which Ctc values could be calculated, 25 (65.8%) had evidence of intraocular production of T gondii-specific antibody on the basis of either an IgM or IgG Ctc value of 1 or greater. The C values and Ctc values were in agreement for 75.9% of IgM containing samples and 75% of IgG containing samples. Sensitivity, specificity, predictive value of a positive test result, and predictive value of a negative test result for an IgM or IgG C value, when compared with the corresponding IgM or IgG Ctc value were determined. The results indicate that use of the C value for estimation of intraocular T gondii-specific antibody production will result in 28.6 (IgM) to 50% IgG) false-negative results and 12.5% (IgM and IgG) false-positive results, when compared with the Ctc value.

Magnification of cervical radiographs prevents accurate interpretation of vertebral canal absolute minimum sagittal diameter (MSD) values and application of the established MSD values for diagnosis of cervical stenotic myelopathy (CSM). Variability in MSD determination in human beings, owing to radiographic magnification, is minimized by assessing a ratio of the vertebral canal diameter to the sagittal width of the vertebral body. This relative measurement technique improves the accuracy of diagnosis of cervical spinal stenosis in human beings. The MSD of the vertebral canal was determined in 50 horses with CSM and 50 control horses, using a radiopaque marker method for correction of magnification. In addition, a ratio of the absolute MSD to the sagittal width of the vertebral body and a ratio of the absolute MSD to the length of the vertebral body were determined in 100 CSM-affected and 100 control horses. Response operating characteristic curve analysis of each method determined that the sagittal ratio method of canal diameter assessment provided the most accurate interpretation of cervical radiographs for diagnosis of CSM, with sensitivity and specificity of larger than or equal to 89% at each vertebral site. The accuracy of the ratio method, without consideration of bony malformation, supports the importance, and perhaps prerequisite, of generalized vertebral canal stenosis in the pathogenesis of CSM. Subjective evaluation of bony malformations from cervical radiographs of 100 CSM-affected horses and 100 control horses indicated that CSM-affected horses have more severe bony malformation than do control horses. However, moderate to marked degenerative joint disease of the articular processes was frequently observed in control horses. Subjective evaluation of bony malformation does not distinguish between CSM-affected and unaffected horses.

A relation exists between colloid osmotic pressure and serum total protein concentration; equations describing this relation have been used to determine a calculated value for colloid osmotic pressure. However, the relation between total protein concentration and colloid osmotic pressure is altered by the method used to measure protein and by changes in the ratio of concentrations of albumin (A) to globulin (G). We developed nomograms for estimating colloid osmotic pressure from A and G concentrations, using samples obtained from clinically normal animals and compared the accuracy of these nomograms with that of previously described equations relating colloid osmotic pressure to total protein concentration. For comparison, serum samples from canine (n = 106), equine (n = 79), feline (n = 24), and bovine (n = 27) patients admitted to the University of Georgia Veterinary Medical Teaching Hospital were used. Results indicated that nomograms based on protein concentration estimated by a refractometer generally were the least reliable. Although predictive nomograms, using total protein concentration determined by the biuret method, provided better results for serum samples, there was considerable variation between measured and calculated values for colloid osmotic pressure in all species studied. Calculated values for colloid osmotic pressure derived from A and G concentrations were most closely related to measured values for colloid osmotic pressure in dogs, horses, and cats. However, calculated values for colloid osmotic pressure differed from measured values by as much as 5 mm of Hg for some samples by each of the methods of estimation. These results indicate that, although calculated values for colloid osmotic pressure may be most accurate when variations in the A-to-G ratio are accounted for in the nomogram, none of the calculation methods provided a consistently accurate estimate of colloid osmotic pressure. For clinical patients, colloid osmotic pressure based on these nomograms cannot replace direct measurement.

Urine uric acid-to-urine creatinine ratios (UUA:UC), urine uric acid concentrations, urine uric acid concentrations corrected for glomerular filtration rate, and urinary uric acid fractional excretions were compared with 24-hour urinary uric acid excretions measured in 6 healthy adult female Beagles. Comparisons, using correlation analysis, were made when dogs consumed a 10.4% protein (dry weight), casein-based diet and a 31.4% protein (dry weight), meat-based diet. The UUA:UC, urine uric acid concentrations corrected for glomerular filtration rate, and urinary uric acid fractional excretions were not reliable estimates of 24-hour urinary uric acid excretions during consumption of either diet. Urine uric acid concentrations in samples collected 2, 4, 6, and 24 hours after initiation of collection correlated with 24-hour urinary uric acid excretions when dogs consumed the casein-based diet; correlation was not found at any time interval when dogs consumed the meat-based diet. Therefore, determination of 24-hour urinary uric acid excretion is recommended because UUA:UC are unreliable.

In veterinary medicine, PCV determined by centrifugation of blood in a microhematocrit tube is the most common clinical test used to initially assess and monitor anemic and polycythemic animals. In contrast, blood hemoglobin (Hb) concentration, rather than PCV, is generally determined in human patients. One automated system photometrically measures blood Hb concentration after conversion of Hb to azide methemoglobin without dilution and was found to be a simple and accurate instrument for use in human medicine. We evaluated the system for its accuracy in measuring blood Hb concentration in animals by comparing it with standard techniques and for its suitability in veterinary practice. Blood samples, anticoagulated with potassium EDTA, from 78 healthy animals (33 dogs, 17 cats, 13 horses, and 15 cows) and 58 dogs and 4 cats with various blood abnormalities (10 anemia, 11 polycythemia, 21 lipemia, 16 leukocytosis, and 6 icterus) were analyzed. In all species, blood Hb concentration of healthy animals determined by the system was comparable to that measured by standard cyanmethemoglobin methods (ie, an automated counter; rI = 0.987 to 0.998 and a hemoglobin kit, rI = 0.946 to 0.993). The aforementioned system also yielded similar values to those obtained by use of standard methods in anemic, polycythemic, and icteric dogs and cats. Moreover, the system reads the absorbance at 2 wavelengths to correct for turbidity, and therefore, accurately measured Hb concentration in blood samples with severe lipemia (triglycerides concentration > 500 mg/dl) and marked leukocytosis (> 50,000 WBC/microl), whereas other standard Hb techniques are known to give falsely high results. We conclude that the automated system compares favorably to standard methods, and is a simple and accurate instrument to quickly measure Hb concentration in animals.

Immunoturbidimetric determination of serum IgG concentration in foals was compared with the reference methods of single radial immunodiffusion and serum protein electrophoresis. High positive correlations were discovered when the technique was compared with either of these reference methods. The zinc sulfate turbidity test for serum IgG estimation was also evaluated. Although a positive correlation was discovered when the latter method was compared with reference methods, it was not as strong as the correlation between reference methods and the immunoturbidimetric method. The immunoturbidimetric method used in this study is specific and precise for equine serum IgG determination. It is rapid and, thus, is advantageous when timely evaluation of critically ill foals is necessary. The technique should be adaptable to various spectrophotometers and microcomputers for widespread application in veterinary medicine.

To determine the suitability and estimate the sensitivity of an immunohistochemical (IHC) test for disease-associated prion protein (PrP(Sc)) in biopsy specimens of rectoanal mucosa-associated lymphoid tissue (RAMALT) for diagnosis of scrapie in sheep. 762 sheep at high risk for having scrapie and indemnified by the National Scrapie Eradication Program. The IHC test for PrP(Sc) was applied to 2 RAMALT and 2 third-eyelid biopsy specimens and a postmortem RAMALT specimen from each sheep. Results were compared with those of a reference test in which results for tissues from obex and retropharyngeal lymph nodes, tonsil, or both were considered in parallel. The reference test identified 139 sheep as having scrapie. Biopsy-related complications occurred in 3 sheep. Sensitivity of the IHC test in RAMALT ranged from 85.3% to 89.4%, depending on the anatomic location from which RAMALT was obtained. Results for the test applied to 1 RAMALT specimen were similar to results interpreted in parallel for 2 third-eyelid specimens (sensitivity, 87.0%). The proportion of inconclusive test results attributable to insufficient lymphoid follicles in biopsy specimens was lower when considering results for 2 RAMALT specimens in parallel (10.1%) than when considering results for 2 third-eyelid specimens in parallel (23.7%). Specimens of RAMALT that were inappropriately collected from an area caudal to the rectoanal interface yielded a high proportion of inconclusive results (33.3% to 50.0%). The IHC test for PrP(Sc) in RAMALT was an effective means of detecting subclinical scrapie in live, high-risk sheep.